Enhancement of fibronectin fibrillogenesis and bone formation by basic fibroblast growth factor via protein kinase C-dependent pathway in rat osteoblasts

Chih Hsin Tang, Rong Sen Yang, Tsang-Hai Huang, Shing Hwa Liu, Wen Mei Fu

Research output: Contribution to journalArticlepeer-review

39 Citations (Scopus)

Abstract

Fibronectin (Fn) is involved in early stages of bone formation and basic fibroblast growth factor (bFGF) is an important factor regulating osteogenesis. We here found that bFGF enhanced extracellular assembly from either endogenously released or exogenously applied soluble Fn in primary cultured osteoblasts. bFGF increased protein levels of Fn using Western blotting analysis. Protein kinase C (PKC) inhibitors such as H7, 3-[1-[3-(amidinothio)propyl-1H-indol-3-yl] -3-(1-methyl-1H-indol-3-yl)maleimide (Bisindolylmaleimide IX), methanesulfonate (Ro 318220,) or 12-(2-cyanoethyl)-6,7,12,13-tetrahydro-13-methyl-5-oxo-5H- indolo[2,3-a]pyrrolo[3,4-c] carbazole (Gö 6976) antagonized the increase of Fn protein by bFGF. Treatment of osteoblasts with bFGF increased membrane translocation of various isoforms of PKC, including α, β, ε, and δ. However, treatment with antisense of various PKC isoforms demonstrated that α and β isozymes play important roles in the enhancement action of bFGF on Fn assembly. Down-regulation of PKC by prolonged treatment with 1 μM 12-O-tetradecanoylphorbol-13 acetate for 24 h inhibited the potentiating action of bFGF. It has been reported that α5β1 integrin is related to Fn fibrillogenesis, and immunocytochemistry showed that bFGF treatment increased the clustering of α5 integrins. Flow cytometry analysis demonstrated that bFGF increased cell surface expression of α5 and β1 integrins and PKC inhibitors antagonized the increase by bFGF. Local administration of bFGF into the metaphysis of the tibia via the implantation of a needle cannula significantly increased the protein levels of Fn in the area of trabecular spongiosa, which was inhibited by coadministration of PKC inhibitors. Furthermore, local injection of bFGF increased the bone volume of secondary spongiosa in tibia, which was significantly antagonized by PKC inhibitors. These results suggest that bFGF increased bone formation and Fn fibrillogenesis both in vitro and in vivo via PKC-dependent pathway.

Original languageEnglish
Pages (from-to)440-449
Number of pages10
JournalMolecular Pharmacology
Volume66
Issue number3
Publication statusPublished - 2004 Sep 1

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Pharmacology

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